Matsubara T, Beeman R W, Shike H, Besansky N J, Mukabayire O, Higgs S, James A A, Burns J C
Department of Pediatrics, University of California, San Diego, School of Medicine, La Jolla, 92093, USA.
Proc Natl Acad Sci U S A. 1996 Jun 11;93(12):6181-5. doi: 10.1073/pnas.93.12.6181.
The lack of efficient mechanisms for stable genetic transformation of medically important insects, such as anopheline mosquitoes, is the single most important impediment to progress in identifying novel control strategies. Currently available techniques for foreign gene expression in insect cells in culture lack the benefit of stable inheritance conferred by integration. To overcome this problem, a new class of pantropic retroviral vectors has been developed in which the amphotropic envelope is completely replaced by the G glycoprotein of vesicular stomatitis virus. The broadened host cell range of these particles allowed successful entry, integration, and expression of heterologous genes in cultured cells of Anopheles gambiae, the principle mosquito vector responsible for the transmission of over 100 million cases of malaria each year. Mosquito cells in culture infected with a pantropic vector expressing hygromycin phosphotransferase from the Drosophila hsp70 promoter were resistant to the antibiotic hygromycin B. Integrated provirus was detected in infected mosquito cell clones grown in selective media. Thus, pantropic retroviral vectors hold promise as a transformation system for mosquitoes in vivo.
缺乏针对重要医学昆虫（如按蚊）的高效稳定遗传转化机制，是阻碍新型控制策略研究进展的最重要因素。目前用于在培养昆虫细胞中表达外源基因的技术，缺乏整合所赋予的稳定遗传优势。为克服这一问题，已开发出一类新型泛嗜性逆转录病毒载体，其中嗜异性包膜被水泡性口炎病毒的G糖蛋白完全取代。这些病毒颗粒拓宽的宿主细胞范围，使得异源基因能够成功进入、整合并在冈比亚按蚊（每年传播超过1亿例疟疾的主要蚊媒）的培养细胞中表达。用来自果蝇hsp70启动子的潮霉素磷酸转移酶表达的泛嗜性载体感染的培养蚊细胞，对潮霉素B具有抗性。在选择性培养基中生长的感染蚊细胞克隆中检测到整合的前病毒。因此，泛嗜性逆转录病毒载体有望成为一种体内转化蚊子的系统。
